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WO2015063069A1 - Récepteurs antigéniques chimériques avec domaines de liaison à l'antigène dérivés des récepteurs des cellules t gamma delta - Google Patents

Récepteurs antigéniques chimériques avec domaines de liaison à l'antigène dérivés des récepteurs des cellules t gamma delta Download PDF

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Publication number
WO2015063069A1
WO2015063069A1 PCT/EP2014/073087 EP2014073087W WO2015063069A1 WO 2015063069 A1 WO2015063069 A1 WO 2015063069A1 EP 2014073087 W EP2014073087 W EP 2014073087W WO 2015063069 A1 WO2015063069 A1 WO 2015063069A1
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amino acid
acid sequence
seq
protein
cells
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PCT/EP2014/073087
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Benjamin Felder
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Benjamin Felder
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/70503—Immunoglobulin superfamily
    • C07K14/7051—T-cell receptor (TcR)-CD3 complex
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/10—Cellular immunotherapy characterised by the cell type used
    • A61K40/11—T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/30—Cellular immunotherapy characterised by the recombinant expression of specific molecules in the cells of the immune system
    • A61K40/31—Chimeric antigen receptors [CAR]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K40/00—Cellular immunotherapy
    • A61K40/40—Cellular immunotherapy characterised by antigens that are targeted or presented by cells of the immune system
    • A61K40/41—Vertebrate antigens
    • A61K40/42—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/03—Fusion polypeptide containing a localisation/targetting motif containing a transmembrane segment

Definitions

  • the present invention relates to engineered membrane proteins comprising: (i) a signal peptide, (ii) an extracellular antigen recognition domain, derived from the variable domains of ⁇ T cell receptor chains (iii) an (optional) spacer region (iv) a transmembrane region and (v) an intracellular effector domain.
  • the present invention furthermore relates to nucleic acids encoding the proteins, expression constructs for expressing the protein in a host cell and host cells.
  • the proteins of the invention are chimeric antigen receptors, that due to their antigen binding region allow to redirect the effector functions of immune cells against a broad spectrum of cancer or virally infected cells.
  • the chimeric antigen receptors of the invention can be used to generate therapeutic cells, suitable for the immunological treatment of malignant or viral disease.
  • the present invention also relates to methods for generating antigen-specific effector cells as well as to methods for the treatment of diseases, in particular cancer, and methods of immunotherapy, preferably including adoptive, target-cell specific immunotherapy.
  • T cell based therapies currently limited due to a variety of factors, including the lack of suitable tumor antigens, the tolerance against tumor antigens, the small repertoire of endogenous tumor antigen specific T cells, and a variety of evasion mechanisms cancer cells employ to prevent their immunological removal (for reviews see Topfer et al., 2011; Capietto et al., 2011). The importance of these limiting factors is exemplified by the unconvincing clinical efficacy of therapeutic cancer vaccines up to date (Huber et al., 2012).
  • Synthetic biology offers new ways to improve the immunologic recognition and elimination of cancer cells.
  • One promising strategy involves the transduction of polyclonal T cells with DNA constructs, encoding engineered membrane receptors that couple the binding of an antigen specific targeting molecule, usually a single-chain variable fragment (scFv), to the delivery of a tailored T cell activating signal.
  • scFv single-chain variable fragment
  • Such artificial T cell receptors due to their composite make-up, commonly referred to as chimeric antigen receptors (CARs), endow their host cells with a predefined antigen specificity (for a review see Sadelain at al., 2013). Thereby the barriers and incremental kinetics of immunization can be bypassed and an immune reaction raised against any surface antigen on target cells in an MHC-independent manner.
  • CAR T cells gain the ability to overcome tumor immune suppression (Loskog et al., 2006). Because T cells are capable of proliferation, the adoptive transfer of tumor-reactive T cells has the additional advantage of an amplification of the anti-tumor response in vivo.
  • the CAR approach is shown to be functionally superior to antibody-dependent cell-mediated cytotoxicity (ADCC), especially within the context of the tumor microenvironment where exogenously introduced anti-tumor antibodies show only weak penetration and short persistence (Boissel et al., 2013). Those key properties single the CAR approach out to provide effective cancer therapeutics.
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • cytotoxic T cells can be grouped into two distinct subpopulations. While 95% of human CD8 cells express a receptor constituted of a and ⁇ subunits, a small proportion express an alternate receptor consisting of ⁇ and ⁇ chains. Short protein fragments presented by MHC complex on the cell surface are the antigens recognized by ⁇ T cells. In contrast, ⁇ T cells possess a restricted T cell receptor diversity, recognizing a variety of non-protein antigens (for a review see Carding and Egan., 2002). One prominent example is the recognition of microbial phosphoantigens like isopentenyl pyrophosphate (IPP) by Vy9/V52 T cells.
  • IPP isopentenyl pyrophosphate
  • IPP is also displayed on the surface of many cancer cells but not normal cells, a circumstance thought to be caused by metabolic aberrations which are common in transformation (Kabelitz et al., 2004). Indeed Marcu-Malina et al. (2011) demonstrated the interesting ability of Vy9/V82 TCR transduced lymphocytes to specifically lyse 17 of 20 cancer cell lines, while untransformed cells remained untouched. This capability of Vy9/V62 T cells to discern a broad spectrum of cancer cells over their healthy counterparts is remarkable in so far that universal cancer-specific antigens otherwise remain at large. Therefore any immunological cancer therapy directed against IPP as an antigen may be suitable for a very large number of patients, including those for whom no effective treatment options exist today.
  • WO 00/31239 describes immune cells having a predefined specificity, wherein the cell is complexed either with an antigen-specific MHC-restricted chimeric T cell receptor or is transfected with an antigen-specific MHC-restricted chimeric TCR gene.
  • WO 2013/147606 describes Vy9/V52 T cell receptor chains with enhanced affinity and target recognition, due to point mutations in their CDR sequences.
  • Zheng et al. (2013) describe soluble TCRs, derived from a fusion of Yy9 and V52 variable domains to an Fc antibody fragment.
  • the fusion protein is shown to couple the ability of the ⁇ -TCR to recognize many cancer cell lines with the ability of the Fc fragment to induce antibody dependent cytotoxicity.
  • the present invention aims to use CAR technology for targeting the IPP antigen, in order to raise potent immune responses against a broad-spectrum of cancer cells.
  • proteins comprising:
  • an antigen recognition domain which is derived from the variable domains of Vy9 V52 T cell receptors (TCRs), (iii) optionally, a spacer region, connecting domain (ii) and domain (iv),
  • nucleic acids encoding the proteins of the present invention.
  • this objective is solved by an expression construct for expressing the protein of the present invention in a cell.
  • this object is solved by a host cell expressing a protein of the present invention or comprising a nucleic acid of the present invention or an expression construct of the present invention.
  • this object is solved by using a protein of the present invention, a nucleic acid of the present invention or an expression construct of the present invention for generating target-specific effector cells.
  • this object is solved by providing the protein of the present invention, the nucleic acid of the present invention, the expression construct of the present invention or the host cell of the present invention for use as a medicament.
  • this object is solved by providing the protein of the present invention, the nucleic acid of the present invention, the expression construct of the present invention or the host cell of the present invention for use in the treatment of cancer or for use in adoptive, target-cell specific immunotherapy.
  • lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, neutrophils, macrophages, mast cells, dendritic cells, killer dendritic cells, B cells;
  • lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, neutrophils, macrophages, mast cells, dendritic cells, killer dendritic cells, B cells;
  • step (c) transferring the protein, nucleic acid, or expression construct provided in step (a) into the host cell or cell line provided in step (b);
  • this object is solved by a method for the treatment of diseases, in particular cancer, comprising the step of
  • a protein of the present invention a nucleic acid of the present invention, an expression construct of the present invention, a host cell of the present invention or an antigen-specific effector cell as obtained according to the method for generating antigen-specific effector cells, and (b) optionally, respective excipient(s).
  • a protein of the present invention a nucleic acid of the present invention, an expression construct of the present invention, a host cell of the present invention or an antigen-specific effector cell as obtained according to the method for generating antigen-specific effector cells, and (b) optionally, respective excipient(s).
  • the present invention provides proteins, in particular chimeric antigen receptors, whose antigen binding domains are derived from endogenous Vy9/V52 TCRs.
  • the proteins of the present invention comprise several domains:
  • TCRs Vy9/V52 T cell receptors
  • the proteins of the invention are preferably cell surface receptor proteins and, thus, comprise:
  • the functionality of the proteins of the invention within a host cell is detectable in an assay suitable for demonstrating the signaling potential of said protein upon binding of a particular ligand. Such assays are available to the skilled artisan.
  • chimeric antigen receptors Upon binding to the target, such chimeric antigen receptors link to endogenous signaling pathways in a cell (an effector cell) and generate certain activating signals (depending on the effector domain).
  • CAR chimeric antigen receptors
  • target specificity such as target-cell specificity
  • NK natural killer
  • CARs generates antigen-specific effector cells for the use in adoptive, target-cell specific immunotherapy.
  • CARs are composed of a target or antigen specific recognition domain or cell recognition domain (domain (ii), such as an scFv antibody fragment) for recognition of an antigen or a target (such as a tumor-cell surface antigen) fused via a flexible spacer region to a transmembrane domain and an effector domain comprising one or more intracellular signaling domains (like the zeta-chain of the CD3 complex of the T- cell receptor).
  • CAR expression retargets the cytotoxic activity of the effector cells (lymphocytes) to antigens/targets (tumor cells) and triggers their cytolysis by the CAR expressing immune effector cells.
  • binding of the antigen / target specific recognition domain of the CAR to its cognate antigen/target on the surface of target cells/viruses transmits a signal into the CAR expressing immune effector cells via the intracellular signaling domain(s) of the CAR which activates the endogenous cytotoxic activity of such immune effector cells.
  • a “signal peptide” refers to a short peptide sequence that destines a protein to a specific location within the cell.
  • the signal peptide of the invention can be any signal peptide of a human secreted or type I transmembrane protein, enabling the correct localization of the CARs of the invention to the cell membrane, in particular the extracellular portion (domains)
  • transmembrane portion (iv) inserted into the plasma membrane and the cytoplasmic portion (v) within the host cell.
  • the signal peptide is cleavable and thereby eliminated before the CAR reaches the self-surface.
  • the signal peptide (i) comprises or is derived from human Alpha- 1 Antitrypsin signal peptide (liAAT) (uniprot accession P01009, amino acid residues 1-24) [SEQ ID NO. 1]:
  • the antigen recognition or binding domain (ii) binds the phosphoantigen IPP on the surface of a target cell.
  • the antigen binding region is understood to be essential for the functionality of the proteins of the invention.
  • antigen recognition domain refers to domain (ii) of the proteins of the invention.
  • the antigen recognition domain is derived from the variable domains of Vy9 V52 T cell receptors (TCRs).
  • TCRs T cell receptors
  • the target is a cell or a virus, more preferably a malignant or a virally infected cell.
  • the antigen recognition domain (ii) comprises or consists of
  • variable domain of Vy9 comprises or consists of an amino acid sequence of SEQ ID NO. 3 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 3.
  • variable domain of V62 comprises or consists of an amino acid sequence of SEQ ID NO. 4 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 4.
  • the antigen recognition domain (ii) comprises the variable domain of Vy9 and the variable domain of V52, which are covalently coupled or fused to each other.
  • variable domain of Vy9 and V62 is via a (flexible) linker (L) yielding a single chain, wherein the variable d domains are either positioned in the sequence of from N- to C-terminus:
  • the orientation is Vy9-L-V52.
  • a preferred linker comprises or consists of the amino acid sequence of SEQ ID NO. 2:
  • the protein of the present invention (in particular the domain (ii) thereof) comprises an amino acid sequence of SEQ ID NO. 5 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 5.
  • the antigen recognition domain (ii) comprises the variable domain of Vy9 and the variable domain of V52, which are not covalently coupled or fused to each other.
  • variable domain of Vy9 and the variable domain of V52 are preferably co-expressed in the host cell.
  • the protein of the present invention comprises two protein chains preferably not covalently coupled to each other and/or each comprising
  • TCRs Vy9 V52 T cell receptors
  • the antigen recognition domain (ii) of one protein chain comprises or consists of the variable domain of Vy9 and the antigen recognition domain (ii) of the other protein chain comprises or consists of the variable domain of V62.
  • the target specific recognition domain serves for the targeting of the protein of the present invention or a respective cell expressing/carrying the protein of the present invention on its surface to a specific target. Binding of the target specific recognition domain of the protein of the present invention (CAR) to its cognate target on the surface of target cells/viruses furthermore transmits a signal into the protein (CAR)-expressing immune effector cells via the intracellular signaling domain(s) of the protein of the present invention which activates the endogenous cytotoxic activity of such immune effector cells.
  • CAR target specific recognition domain of the protein of the present invention
  • a CAR comprising Vy9 (a) and a CAR comprising V62 can be co-expressed within the same host cell.
  • variable domains are mutated at key residues to enhance their stability, this might allow to omit T cell receptor constant regions as a stabilizing element.
  • the antigen recognition domain (ii) comprises one or more CDR sequences derived from the variable domains of Vy9 and/or V62, or an amino acid sequence that has at least 70%, preferably 80% or more preferably 90% or 95% sequence identity to said sequences.
  • the CDR sequences comprise or consist of
  • the antigen recognition domain (ii) may comprise or consist of CDR sequences derived from the variable domains of Vy9 and/or V52 grafted onto protein scaffolds. These scaffolds can be distinct from the native variable domains the CDR sequences are derived from. Any protein which allows to exchange certain unstructured portions, while maintaining an overall stable tertiary structure may be suitable as a scaffold. Examples of such scaffolds which might be suitable to accept CDR sequences as grafts are variable domains of immunoglobulin or the extracellular type III domains of fibronectin.
  • the spacer region (iii) connects the antigen recognition domain (ii) and the transmembrane domain (iv).
  • the spacer region serves as a flexible link between the antigen recognition domain (ii) and the transmembrane domain (iv) and the effector domain (v). It ensures the necessary accessibility and flexibility of the antigen recognition domain (ii).
  • the spacer region (iii) comprises or consists of the constant domains of any TCR chain, in order to enhance the stability and surface expression of the variable domains comprises in the antigen recognition domain (ii).
  • the spacer region (iii) comprises constant domain(s) of a T cell receptor.
  • said constant domain(s) of a T cell receptor are selected from Cy, C5, Ca and/or Cp.
  • the constant domain Gy comprises an amino acid sequence of SEQ ID NO. 6 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 6;
  • the constant domain C6 comprises an amino acid sequence of SEQ ID NO. 7 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 7;
  • the constant domain Ca comprises an amino acid sequence of SEQ ID NO. 8 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 8; and/or
  • the constant domain ⁇ comprises an amino acid sequence of SEQ ID NO. 9 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 9.
  • a transmembrane domain comprises a stretch of hydrophobic amino acids which by virtue of its hydrophobicity can thermodynamically stable integrate into the cell membrane.
  • the transmembrane domain (iv) within the protein of the invention anchors it to the cell membrane, whereby antigen binding (ii) and the spacer domains (iii) are positioned extracellularly and the effector domain (v) on the cytoplasmatic site.
  • the transmembrane domain (iv) comprises or consists of the transmembrane domain of human CD28, preferably comprising an amino acid sequence of SEQ ID NO. 10 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to the amino acid sequence of SEQ ID NO. 10. human CD28.
  • the effector domain (v) connects to the C-terminus of the transmembrane domain (iv).
  • the effector domain (v) comprises one or more intracellular signaling domains and activates the intracellular signalling machinery upon antigen binding, activating the cytotoxic/effector function of the host cell and/or promoting its proliferation and/or cytokine secretion.
  • the at least one effector domain (v) of the protein of the invention preferably comprises endodomains/domain(s) of at least one of the following:
  • CD3 polypeptides ⁇ , ⁇ , ⁇
  • FceRIy B29
  • CD28 such as CD28, OX40 (CD134), 4-1BB (CD137).
  • At least one effector domain (v) that comprises or consists of (is):
  • the LL motif of the CD28 fragment is inactivated by mutation (such as to a diglycine motif, other functional similar mutations are possible as well) to enhance protein stability and surface of expression of the protein of the invention.
  • the respective CD28 fragment preferably comprises or consists of the amino acid sequence of SEQ ID NO. 13, or a functional equivalent thereof;
  • the term "functional equivalent” defines a protein or nucleotide sequence, having a different amino acid or base sequence, compared to the sequences disclosed herein, but exhibiting the same function in vitro and in vivo.
  • An example of a functional equivalent is a modified or synthetic gene, encoding the expression of a protein identical or highly homologous to that encoded by the wildtype gene or a sequence disclosed herein.
  • the effector domain (v) (preferably) comprises or consists of (is) an amino acid sequence with the amino acid sequence of SEQ ID NO. 15 or a functional equivalent thereof, wherein a "functional equivalent” has less sequence identity (such as at least 80% sequence identity, preferably at least 90% sequence identity, more preferably at least 95% sequence identity or 99% sequence identity) but is a functional fusion of the costimulatory CD28 receptor (diglycine motif mutated) fused to a fragment of the zeta-chain of the CD3 complex of the T- cell receptor fused to a fragment of the costimulatory OX40 receptor.
  • a "functional equivalent” has less sequence identity (such as at least 80% sequence identity, preferably at least 90% sequence identity, more preferably at least 95% sequence identity or 99% sequence identity) but is a functional fusion of the costimulatory CD28 receptor (diglycine motif mutated) fused to a fragment of the zeta-chain of the CD3 complex of the T- cell receptor fused to a fragment
  • the according to the invention comprises or consists of the amino acid sequence of:
  • an antigen binding domain that comprises or consists of (is) a variable domain of a Vy9-L-V82 (SEQ ID NO. 5)
  • a spacer region preferably comprising a constant region of a T cell receptor (SEQ ID NOs. 6 to 9)
  • transmembrane domain as defined herein, preferably of SEQ ID NO. 10
  • the protein comprises or consists of the amino acid sequence of SEQ ID NO. 16; or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to SEQ ID NO. 16.
  • amino acid sequence of SEQ ID NO. 16 refers to the amino acid sequence of the multifunctional protein with the domains:
  • the protein comprises or consists of the amino acid sequence of SEQ ID NO. 17; or an amino acid sequence that has at least 70%) sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to SEQ ID NO. 17.
  • amino acid sequence of SEQ ID NO. 17 refers to the amino acid sequence of the multifunctional protein with the domains:
  • the protein comprises or consists of the amino acid sequence of SEQ ID NO. 18; or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% sequence identity to SEQ ID NO. 18.
  • amino acid sequence of SEQ ID NO. 18 refers to the amino acid sequence of the multifunctional protein with the domains:
  • the protein of the present invention comprises or consists of the amino acid sequence of any of SEQ ID NOs. 16 to 18 or an amino acid sequence that has at least 70% sequence identity, preferably 80%, more preferably 90% or more preferably 95% or 99% sequence identity to the amino acid sequence of any of SEQ ID NOs. 16 to 18.
  • the present invention provides nucleic acids/nucleic acid molecules/isolated nucleic acid molecules encoding the proteins of the invention.
  • the nucleic acids according to this invention comprise DNA (such as dsDNA, ssDNA, cDNA), RNA (such as dsRNA, ssRNA, mRNA), combinations thereof or derivatives (such as PNA) thereof.
  • a nucleic acid of the invention comprises or consists of the nucleic acid encoding for the amino acid sequence of SEQ ID NO. 16,
  • nucleic acid sequence of SEQ ID NO. 19 or comprises or consists of the nucleic acid sequence of SEQ ID NO. 19 or their complementary sequences or sequences that have at least 95 % sequence identity.
  • a nucleic acid of the invention comprises or consists of the nucleic acid encoding for the amino acid sequence of SEQ ID NO. 17,
  • nucleic acid sequence of SEQ ID NO. 20 or comprises or consists of the nucleic acid sequence of SEQ ID NO. 20 or their complementary sequences or sequences that have at least 95 % sequence identity.
  • a nucleic acid of the invention comprises or consists of the nucleic acid encoding for the amino acid sequence of SEQ ID NO. 18,
  • nucleic acid sequence of SEQ ID NO. 21 or their complementary sequences or sequences that have at least 95 % sequence identity.
  • nucleic acids comprising or consisting of nucleic acid sequences of SEQ ID NOs. 16 and 17 (or respective sequences with at least 90% identity) are for co-expression in a host cell (to yield a functional receptor/CAR).
  • the nucleic acid sequences of the present invention are codon-optimized for expression in mammalian cells, preferably for expression in human cells.
  • Codon-optimization refers to the exchange in a sequence of interest of codons that are generally rare in highly expressed genes of a given species by codons that are generally frequent in highly expressed genes of such species,- such codons encoding the same amino acids as the codons that are being exchanged.
  • nucleotide sequences obtained due to the degeneration of the genetic code of the above nucleotide sequences are also the nucleotide sequences obtained due to the degeneration of the genetic code of the above nucleotide sequences.
  • the present invention provides expression constructs for expressing the protein of the invention in a cell.
  • the expression constructs further comprise promoter and terminator sequences.
  • An "expression construct” or “gene construct” refers to a nucleic acid construct, usually an expression vector or plasmid, that is used to introduce a specific gene sequence into a target cell. Once the expression or gene construct is inside the cell, the protein that is encoded by the gene is produced by the cellular transcription and translation macliinery.
  • the expression or gene construct is designed to contain respective regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene earned on the construct, including promoter and terminator sequences. The goal of a well-designed expression or gene construct is the production of suitable amounts of stable mRNA, and therefore proteins.
  • nucleic acids and/or in particulai- expression constructs of the invention are capable of directing the synthesis/expression of the engineered membrane proteins of the invention.
  • nucleic acids and/or expression constructs of the invention are dsDNA, ssD A, RNA or mRNA or combinations thereof.
  • the present invention provides host cells which express a protein of the invention or which comprise a nucleic acid or an expression construct of the invention.
  • the host cell is selected from effector cells of the immune system, such as lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, neutrophils, macrophages, dendritic cells, killer dendritic cells, B cells.
  • lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, mast cells, neutrophils, macrophages, dendritic cells, killer dendritic cells, B cells.
  • Effective cells of the immune system or “immune effector cells” refers to cells of hematopoietic origin including but not limited to the cell types mentioned above that are functionally involved in the initiation and/or execution of innate and/or adaptive immune responses.
  • the protein of the invention comprises only the variable domain Vy9 or V52 as antigen binding domain (ii),
  • the host cell co-expresses two proteins of the invention or comprises two nucleic acids or expression constructs of the invention
  • first protein comprising the variable domain Vy9 as antigen binding domain (ii) and a second protein comprising the variable domain V82 as antigen binding domain (ii), respective nucleic acids/expression constructs of said two proteins,
  • proteins comprising or consisting of the amino acid sequences of SEQ ID NOs. 16 and 17 (or respective sequences with at least 90% identity), or nucleic acids comprising or consisting of the nucleic acid sequences of SEQ ID NOs. 19 and 20 (or respective sequences with at least 90% identity).
  • the invention provides the use of the protein, nucleic acid, or expression construct of the present invention for generating antigen-specific effector cells.
  • Antigen-specific effector cells or “target-specific effector cells” refer to effector cells of the immune system or immune effector cells genetically modified to express the multi-functional protein of the invention by transfer of an expression construct or nucleic acid encoding said multi-functional protein.
  • Such antigen-specific or target-specific effector cells are versatile means, in particular in the treatment of diseases (as described below for ACT and cancer treatment).
  • the invention provides the protein, nucleic acid, expression construct or host cell for use as a medicament.
  • the invention provides the protein, nucleic acid, expression construct or host cell for use in the treatment of cancer.
  • the invention provides the protein, nucleic acid, expression construct or host cell for use in adoptive, target-cell specific immunotherapy.
  • “Adoptive, target-cell specific immunotherapy” refers to a form of therapy in which immune cells are transferred to tumor-bearing hosts.
  • the immune cells have antitumor reactivity and can mediate direct or indirect antitumor effects.
  • ACT adoptive cell therapy
  • immune effector cells such as lymphocytes with anti-tumour activity
  • lymphocytes with anti-tumour activity expanded in vitro and infused into the patient with cancer.
  • ACT using autologous tumour- infiltrating lymphocytes has emerged as the most effective treatment for patients with metastatic melanoma and can mediate objective cancer regression in approximately 50% of patients.
  • the use of donor lymphocytes for ACT is an effective treatment for immunosuppressed patients who develop post-transplant lymphomas (reviewed in Rosenberg et al., 2008).
  • the invention provides methods for generating antigen-specific effector cells.
  • lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, neutrophils, macrophages, mast cells, dendritic cells, killer dendritic cells, B cells;
  • lymphocytes including but not limited to cytotoxic lymphocytes, T cells, cytotoxic T cells, T helper cells, Thl7 T cells, natural killer (NK) cells, natural killer T (NKT) cells, neutrophils, macrophages, mast cells, dendritic cells, killer dendritic cells, B cells;
  • step (c) transferring the protein, nucleic acid, or expression construct provided in step (a) into the host cell or cell line provided in step (b);
  • the present invention also provides methods for the treatment of diseases, in particular cancer, and methods of immunotherapy, preferably including adoptive, target-cell specific immunotherapy.
  • the method for the treatment of diseases, in particular cancer comprises the step of administering to a subject in need thereof a therapeutically effective amount of (a) a protein, a nucleic acid, an expression construct or a host cell (in particular an antigen-specific effector cell) as obtained and defined herein, and (b) optionally, respective excipient(s).
  • the method of immunotherapy preferably including or utilizing adoptive, target-cell specific immunotherapy, according to the present invention comprises the step of administering to a subject in need thereof a therapeutically effective amount of (a) a protein, a nucleic acid, an expression construct or a host cell (in particular an antigen-specific effector cell) as obtained and defined herein, and (b) optionally, respective excipient(s).
  • a therapeutically effective amount of a protein, a nucleic acid, an expression construct or a host cell (in particular an antigen-specific effector cell) of this invention refers to the amount that is sufficient to treat the respective disease or achieve the respective outcome of the adoptive, target-cell specific immunotherapy.
  • FIG. 1 Schematic structure of the expression vectors for single-chain constructs.
  • V - promoter SP - signal peptide
  • V - variable domain Vy9 or V52
  • L - linker C - constant domain
  • TM - transmembrane domain TM - transmembrane domain
  • ED - effector domain pA - polyadenylation signal (terminator)
  • ABR - antigen binding region V - promoter, SP - signal peptide, V - variable domain (Vy9 or V52), L - linker, C - constant domain, TM - transmembrane domain, ED - effector domain, pA - polyadenylation signal (terminator), ABR - antigen binding region
  • Figure 2 Dotplot representation of flow -cytometric CAR expression analysis.
  • NC-E - only T cells NC-T only target cell
  • mock Vy9/V52 coexpression of both Yj9 and
  • V52 comprising CARs, sc_Vy9V62-expression of CAR with fusion ABR
  • Vy9, V52 and single-chain Vy9V62 mock CARs, codon optimized for human cells, were synthesized by Life Teclmologies (GeneArt® StringsTM DNA Fragments).
  • V62 eIF4G promoter- 5'-UTR-V52-C6-CD28 TM-CD28-CD3zeta-OX40-3'UTR-pA
  • SC_VY9V52 eIF4G promoter- 5*-UTR-SP-VY9-L-V52-C6-CD28 TM-CD28-CD3zeta-OX40- 3'UTR-pA (Fig. 1 B).
  • the mock construct omitted the antigen binding region, yielding the construct:
  • Linear DNA templates were circularized via digestion EcoRI (Biozym, Germany) and subsequent incubation with T4 DNA ligase (Biozym, Germany).
  • 10 ng of circular template was denatured at 95° C for 3 min and cooled to room temperature.
  • the template solution was then added to 1 ml of amplification solution containing phi29 DNA polymerase (10 U, Biozym, Germany), 0,2 mM dNTPs, 100 ig BSA in 50 mM Tri-HCl pH 7,5, 10 mM MgCl, 10 mM (NH) SO, DTT.
  • T cells were transduced with the amplified nucleic acid mock, Vy9, V62, dual Vy9/V2 and the single-chain construct (sc_Vy9V82).
  • sc_Vy9V82 the amplified nucleic acid mock, Vy9, V62, dual Vy9/V2 and the single-chain construct (sc_Vy9V82).
  • phosphate buffered 1 ml Leibovitz LI 5 medium Sigma- Aldrich, Germany
  • 100 g/ml ampicillin Sigma-Aldrich, Germany.
  • GenePORTER 2 transfection reagent BioCat, Heidelberg, Germany was used for cell transfection as directed by the manufacturer.
  • Target cells were provided in 96 well plates with 100,000 cells per well in Leibovitz L15 medium (150 ⁇ ), containing pamidronate (10 ⁇ , Sigma-Aldrich, Germany) and ampicillin (100 ⁇ g/ml).
  • the cytotoxicity of Vy9/V62 is generally higher than that of sc_Vy9 V62, potentially owing to a more native protein configuration. No cytotoxicity of Vy9/V62 and the sc_Vy9V52 can be detected against untransformed PBMC and CD 19+ cells.
  • SEQ ID NO. 1 shows the amino acid sequence of the signal peptide of human Alpha- 1 Antitrypsin
  • SEQ ID NO. 2 shows the amino acid sequence of the preferred linker sequence.
  • SEQ ID NO. 3 shows the amino acid sequence of the variable domain human Vy9 T cell receptor chain.
  • SEQ ID NO. 4 shows the amino acid sequence of the variable domain human V52 T cell receptor chain.
  • SEQ ID NO. 5 shows the amino acid sequence of the fusion of Vy9 to V62, via the linker described under SEQ ID NO. 2.
  • SEQ ID NO. 6 shows the amino acid sequence of the constant domain of the human ⁇ T cell receptor chain.
  • SEQ ID NO. 7 shows the amino acid sequence of the constant domain of the human ⁇ T cell receptor chain.
  • SEQ ID NO. 8 shows the amino acid sequence of the constant domain of the human a T cell receptor chain (uniprot accession number: P01848).
  • SEQ ID NO. 9 shows the amino acid sequence of the constant domain of the human ⁇ T cell receptor chain (uniprot accession number: P01850).
  • SEQ ID NO. 10 shows the amino acid sequence of the transmembrane domain of human CD28 (uniprot accession number: P01850, amino acid residues: 153-179).
  • SEQ ID NO. 11 shows the amino acid sequence of the intracellular domain of human T-cell surface glycoprotein CD3 zeta chain (UniProt accession number P20963-1 (CD3Z_HUMAN); Isoform 1, (amino acid residues 52-164)).
  • SEQ ID NO. 12 shows the amino acid sequence of the intracellular domain of human CD28 costimulatory receptor (UniProt accession number PI 0747-1 (CD28_HUMAN), isoform 1, (amino acid residues 180-220)).
  • SEQ ID NO. 13 shows the amino acid sequence described under SEQ ID NO. 12 wherein the diglycine motif has been inactivated by mutation.
  • SEQ ID NO. 14 shows the amino acid sequence of the intracellular domain of the human costimulatory OX40 receptor (amino acid residues 236-277]) (UniProt accession number P43489 (TN-FRSF4JHUMAN)).
  • SEQ ID NO. 15 shows the amino acid sequence of the effector domain comprising a fusion of CD28 (SEQ ID NO. 13), CD3 zeta (SEQ ID NO. 11) and OX40 (SEQ ID NO. 14), in the mentioned order.
  • SEQ ID NO. 16 shows the amino acid sequence of the multifunctional protein with the domains:
  • SEQ ID NO. 17 shows the amino acid sequence of the multifunctional protein with the domains:
  • SEQ ED NO. 18 shows the amino acid sequence of the multifunctional protein with the domains:
  • SEQ ID NO. 19 shows the nucleotide sequence encoding for the multi-functional protein with the domains:
  • SEQ ID NO. 20 shows the nucleotide sequence encoding for the multi-functional protein with the domains:
  • SEQ ID NO. 21 shows the nucleotide sequence encoding for the multi-functional protein with the domains:
  • SEQ ID NO. 22 shows the amino acid sequence of the CDR3 region of Vy9.
  • SEQ ID NO. 23 shows the amino acid sequence of the CDR3 region of V52.

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Abstract

La présente invention concerne des protéines de membrane obtenues par ingénierie comprenant: (i) un peptide signal, (ii) un domaine de reconnaissance d'antigène extracellulaire, dérivé des domaines variables des chaînes de récepteur des cellules T γδ, (iii) une région espaceur (facultatif), (iv) une région transmembranaire et (v) un domaine effecteur intracellulaire. La présente invention concerne en plus des acides nucléiques codant les protéines, des constructions d'expression pour exprimer la protéine dans une cellule hôte et des cellules hôtes. Les protéines de l'invention sont des récepteurs antigéniques chimériques, qui en raison de leur région de liaison à l'antigène permettent de rediriger les fonctions effectrices des cellules immunitaires contre un large spectre de cellules cancéreuses ou infectées par voie virale. En conséquence, les récepteurs antigéniques chimériques de l'invention peuvent être utilisés pour générer des cellules thérapeutiques, appropriées pour le traitement immunologique de maladies malignes ou virales. La présente invention concerne aussi des procédés pour générer des cellules effectrices spécifiques de l'antigène ainsi que des procédés pour le traitement de maladies, en particulier, le cancer, et des procédés d'immunothérapie, de préférence incluant l'immunothérapie adoptive spécifique des cellules cibles.
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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016201394A1 (fr) * 2015-06-12 2016-12-15 Miltenyi Biotec Technology, Inc. Procédé de traitement de cancer à l'aide de lymphocytes t génétiquement modifiés
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WO2017212072A1 (fr) * 2016-06-10 2017-12-14 Umc Utrecht Holding B.V. Récepteurs de lymphocytes t gamma delta restreints à l'antigène leucocytaire humain et leurs méthodes d'utilisation
KR20170142995A (ko) * 2015-08-05 2017-12-28 주식회사 유영제약 키메라 항원 수용체 및 키메라 항원 수용체가 발현된 t 세포
US10098951B2 (en) 2015-10-23 2018-10-16 Eureka Therapeutics, Inc. Antibody/T-cell receptor chimeric constructs and uses thereof
US10324083B2 (en) 2012-03-28 2019-06-18 Gadeta B.V. Methods of treating cancer in a subject by administering a composition comprising gamma 9 delta 2 T-cell receptors
US10415017B2 (en) 2017-05-17 2019-09-17 Thunder Biotech, Inc. Transgenic macrophages, chimeric antigen receptors, and associated methods
US10822413B2 (en) 2017-04-26 2020-11-03 Eureka Therapeutics, Inc. Cells expressing chimeric activating receptors and chimeric stimulating receptors and uses thereof
US11052138B2 (en) 2015-05-20 2021-07-06 Thunder Biotech Inc. Use of car and bite technology coupled with an SCFV from an antibody against human thymidine kinase 1 to specifically target tumors
CN113728095A (zh) * 2019-04-22 2021-11-30 南京传奇生物科技有限公司 工程化的细胞及其用途
US11352439B2 (en) 2015-08-13 2022-06-07 Kim Leslie O'Neill Macrophage CAR (MOTO-CAR) in immunotherapy
CN114685684A (zh) * 2020-12-31 2022-07-01 博生吉医药科技(苏州)有限公司 MUC1-Tn嵌合抗原受体修饰的Vγ9Vδ2T细胞及其应用
EP4010082B1 (fr) * 2020-08-14 2023-01-25 GammaDelta Therapeutics Limited Anticorps anti-domaine variable 1 de tcr delta multispécifiques
US12161669B2 (en) 2017-05-18 2024-12-10 Umc Utrecht Holding B.V. Compositions and methods for cell targeting therapies

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013147606A1 (fr) * 2012-03-28 2013-10-03 Umc Utrecht Holding B.V. Echange combinatoire de chaîne des récepteurs des cellules γ9δ2t

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2013147606A1 (fr) * 2012-03-28 2013-10-03 Umc Utrecht Holding B.V. Echange combinatoire de chaîne des récepteurs des cellules γ9δ2t

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BERNHARD MOSER ET AL: "Î Î' T-APCs: a novel tool for immunotherapy?", CMLS CELLULAR AND MOLECULAR LIFE SCIENCES, BIRKHÄUSER-VERLAG, BA, vol. 68, no. 14, 15 May 2011 (2011-05-15), pages 2443 - 2452, XP019919462, ISSN: 1420-9071, DOI: 10.1007/S00018-011-0706-6 *
CHIARA RIGANTI ET AL: "Human [gamma][delta] T-cell responses in infection and immunotherapy: Common mechanisms, common mediators?", EUROPEAN JOURNAL OF IMMUNOLOGY, vol. 42, no. 7, 1 July 2012 (2012-07-01), pages 1668 - 1676, XP055168326, ISSN: 0014-2980, DOI: 10.1002/eji.201242492 *

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